Absolute Characterization of Glycoproteins and their Interactions with Proteins and Antibodies by Light Scattering

06 Sep 2017

Glycoproteins (GPs) are primary targets of neutralizing and therapeutic antibodies for a large variety of indications, including epilepsy and many types of viruses. Many non-IgG therapeutic proteins are themselves GPs. Hence, the characterization of GPs and potential degradants, as well as their interactions with therapeutic or target proteins, is essential for understanding GP targets and developing effective therapeutics. This webinar presents two first-principles techniques for carrying out these analyses in solution: SEC-MALS-UV-RI and CG-MALS.
Unlike common unmodified proteins that are globular in conformation and highly hydrophilic in surface chemistry, GPs cannot be characterized accurately by standard analytical size-exclusion chromatography (SEC). Coupling SEC to multi-angle light scattering (MALS) and differential refractive index (dRI) detection results in a powerful SEC-MALS-UV-RI triple-detection system that can determine the molar mass of the protein and total glycan mass in order to characterize degradants and complexes. Composition-gradient multi-angle light scattering (CG-MALS) utilizes the same detectors, with a stop-flow system to quantify the absolute stoichiometry and binding affinity of reversible association between GPs and antibodies or solubilized target proteins.
Join this webinar to learn about:
The limitations of analytical SEC and native PAGE (polyacrylamide gel electrophoresis) for characterizing glycoproteins
How SEC-MALS characterizes glycoproteins, aggregates and complexes
How CG-MALS analyzes self- and hetero-association in solution, intuitively and from first principles
How CG-MALS can be combined with cryo-EM (cryo-electron microscopy) to characterize complex GP-antibody interactions